Method for making a biofabricated material containing collagen fibrils
a biofabricated material and collagen technology, applied in the field of biofabricated leather materials, can solve the problems of increasing the cost of leather, and requiring enormous amounts of feed, etc., and achieves the effects of uniform texture, uniform elasticity or tensile strength, and convenient uniform uptake of dyes and coatings
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example 1
Controlling the Thickness of Biofabricated Leather
[0245]Hydrogels of extacted bovine type I collagen were formed at different collagen concentrations and volumes to produce dried collagen materials of different thicknesses. Collagen was dissolved in 0.01N HCl at either 5 g / L or 9 g / L, then 1 part 10× PBS was added to 9 parts dissolved collagen to induce collagen fibrillation and gel formation.
[0246]Solutions of either 0.8 L or 1.6 L of the fibrillating collagen were then cast into molds and incubated at 25° C. to allow hydrogel formation. The 0.8 L solution produced a gel of 1.5 cm thickness while the 1. 7 L solution produced a gel of 3.0 cm thickness. These gels were dehydrated and lubricated in acetone, then dried and mechanically staked into a leather like material. The thickness of the final dried material correlated with the total amount of collagen in the starting hydrogel.
[0247]The thickness of biofabricated leather was controlled by varying its total collagen content. Sample...
example 2
Production of Biofabricated Leather from Type I Collagen
[0248]Type I collagen was purchased from Wuxi Biot Bio-technology Company, ltd. (Medical Collagen Sponge). The collagen was isolated from bovine tendon by acid treatment followed by pepsin digestion, and was purified by size exclusion chromatography, frozen and lyophilized.
[0249]The lyophilized protein (4.1 g) was dissolved in 733 ml 0.01 N HCL using an overhead mixer. After the collagen was adequately dissolved, as evidenced by a lack of solid collagen sponge in the solution (at least lhr mixing at 1600 rpm), 82 uL of the tanning agent Relugan GTW was added to the solution followed by 81 mL of a 10× PBS, pH 11.2 to raise the pH of the solution to 7.2.
[0250]The solution was then mixed for 3 min before pouring the solution into a silicon mold. The collagen solution was incubated in the silicon mold for 2 hrs at 25° C. to allow the collagen to fibrillate into a viscoelastic hydrogel.
[0251]Plateau of rheological properties along w...
example 3
Production of Biofabricated Leather from Type III Collagen
[0256]A solution of recombinant collagen type III at 2.5 mg / ml in 0.01 N HCl (FibroGen, Inc.) was fibrillated by adding 1 part of a 200 mM of sodium phosphate solution (22 mL), pH 11.2 to 9 parts of the collagen solution (200 mL) to increase the pH to 7 and stirred 2 hours at room temperature.
[0257]Fibrillation was confirmed by measuring 400nm absorbance of the solution over time.
[0258]After fibrillation, the fibrils were tanned by adding Relugan GTW (2% w / w offer on the collagen) to the fibril suspension and mixing for 30 min.
[0259]The tanned collagen fibrils were then centrifuged at 3,500 RPM for 30 minutes to concentrate the fibrils to a concentration of 10 mg / ml . The 10 mg / ml fibril pellet was further centrifuged using an ultra-centrifuge at 21,000 RPM for 30 minutes yielding a fibril gel with a concentration of ˜40-50 mg / ml .
[0260]The physical properties of the fibril gel were assessed with a rheometer.
[0261]The storage...
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